National Repository of Grey Literature 3 records found  Search took 0.01 seconds. 
The monitoring of intracellular ion concentrations in microbial cells
Vodáková, Adéla ; Plášek, Jaromír (advisor) ; Krůšek, Jan (referee)
The Master Thesis focuses on monitoring of intracellular ion concentrations in bacteria Escherichia coli and yeast Saccharomyces cerevisiae using genetically encoded fluorescent probes with green fluorescent protein (GFP). Aquired knowledge about this protein and its spectral characteristics is summarized in the introduction. For experimental study a pH-sensitive sensor which displays a ratio change of two excitation fluorescence peaks - pHluorin - was chosen. This probe was tested in bacteria and yeast cells. The experiments concentrated on the ability of the cell to maintain a constant cytosolic pH under various conditions like different pH values of the suspension, addition of glucose or KCl to the suspension. Another topic discussed in the thesis is the elimination of the cell autofluorescence from the GFP signal. For this purpose the synchronous fluorescence scan technique was succesfully used. I have found out that by using this method the measurements of cytosolic pH values are even more accurate thanks to the improved signal to noise ratio.
The monitoring of intracellular ion concentrations in microbial cells
Vodáková, Adéla ; Plášek, Jaromír (advisor) ; Krůšek, Jan (referee)
The Master Thesis focuses on monitoring of intracellular ion concentrations in bacteria Escherichia coli and yeast Saccharomyces cerevisiae using genetically encoded fluorescent probes with green fluorescent protein (GFP). Aquired knowledge about this protein and its spectral characteristics is summarized in the introduction. For experimental study a pH-sensitive sensor which displays a ratio change of two excitation fluorescence peaks - pHluorin - was chosen. This probe was tested in bacteria and yeast cells. The experiments concentrated on the ability of the cell to maintain a constant cytosolic pH under various conditions like different pH values of the suspension, addition of glucose or KCl to the suspension. Another topic discussed in the thesis is the elimination of the cell autofluorescence from the GFP signal. For this purpose the synchronous fluorescence scan technique was succesfully used. I have found out that by using this method the measurements of cytosolic pH values are even more accurate thanks to the improved signal to noise ratio.
Study of liposomes by using of drop coating deposition Raman spectroscopy
Vodáková, Adéla ; Kočišová, Eva (advisor) ; Bednárová, Lucie (referee)
in english: Goal of the Bachelor Thesis was an optimization of liposome membrane preparation with different composition for the drop coating deposition Raman (DCDR) method, measuring of the DCDR spectra from dried droplets and their interpretation. The DCDR method lies in deposition of small volume of suspension (~μl) on special surface and measuring of Raman spectra from the ring formed at the edge of the dried droplet. The main advantage of used method is the possibility of measuring spectra from the sample with low concentration. Our results show that the DCDR method is very useful in studies of biological membranes with different composition. Two synthetic lipids and one natural extract were chosen for making a comparison. We have found out that spectra obtained from the ring of dried droplet are not different from the spectra measured in the suspension. The important characteristic of spectra is that dried droplet keeps the same lipid phase as in suspension. Spectral mapping proved good reproducibility of the signal inside the ring.

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